Development of the Handy Bio-Strand and its application to genotyping of OPRM1 (A118G)

Anal Biochem. 2007 Aug 1;367(1):79-86. doi: 10.1016/j.ab.2007.04.052. Epub 2007 May 6.

Abstract

We previously developed a three-dimensional microarray system, the Bio-Strand, which exhibits advantages in automated DNA analysis in combination with our Magtration Technology. In the current study, we have developed a compact system for the Bio-Strand, the Handy Bio-Strand, which consists of several tools for the preparation of Bio-Strand Tip, hybridization, and detection. Using the Handy Bio-Strand, we performed single nucleotide polymorphism (SNP) genotyping of OPRM1 (A118G) by allele-specific oligonucleotide competitive hybridization (ASOCH). DNA fragments containing SNP sites were amplified from genomic DNA by PCR and then were fixed on a microporous nylon thread. Thus, prepared Bio-Strand Tip was hybridized with allele-specific Cy5 probes (<15mer), on which the SNP site was designed to be located in the center. By optimizing the amount of competitors, the selectivity of Cy5 probes increased without a drastic signal decrease. OPRM1 (A118G) genotypes of 23 human genomes prepared from whole blood samples were determined by ASOCH using the Handy Bio-Strand. The results were perfectly consistent with those determined by PCR direct sequencing. ASOCH using the Handy Bio-Strand would be a very simple and reliable method for SNP genotyping for small laboratories and hospitals.

Publication types

  • Evaluation Study

MeSH terms

  • Alleles
  • Base Sequence
  • DNA / genetics
  • DNA / isolation & purification
  • Equipment Design
  • Genotype
  • Humans
  • Oligonucleotide Array Sequence Analysis / instrumentation*
  • Oligonucleotide Array Sequence Analysis / methods
  • Oligonucleotide Probes / genetics
  • Polymorphism, Single Nucleotide*
  • Receptors, Opioid, mu / genetics*

Substances

  • OPRM1 protein, human
  • Oligonucleotide Probes
  • Receptors, Opioid, mu
  • DNA